Quickgene dna whole blood kit

DNA was extracted from the peripheral blood of 231 patients who had kidney transplantation between January 1996 and December 2004 by using the LaboPass Genomic DNA Extraction Kit (COSMO, Seoul, Korea) or QuickGene DNA whole blood kit (Fujifilm, Tokyo, Japan) when HLA genotyping for pre-operation evaluation was performed. For 195 healthy controls, DNA was extracted using LaboPass Genomic DNA Extraction Kit (COSMO, Seoul, Korea) during the period of January 1999 and July 2001. All DNA samples were preserved at –70℃ prior to being used for Foxp3 polymorphisms analyses which were performed during the period of June 2015 and July 2016. Four Foxp3 polymorphisms (rs3761548 A/C, rs2280883 C/T, rs5902434 del/ATT, and rs2232365 A/G) were analyzed by PCR with sequence-specific primers (PCR-SSP) with some modifications [20 (link)] (Table 2). PCR was performed by using a 40-µL reaction mixture containing 40 ng DNA, 0.2mM of each primer, 0.8 µL of 10mM dNTP, 2.0mM MgCl₂, 1.0 U Taq DNA polymerase (Roche Applied Science, Basel, Switzerland), and 4 µL of 10× reaction buffer. The PCR protocol consisted of an initial denaturation step at 95℃ for 5 min; 35 cycles of denaturation at 95℃ for 30 sec, annealing (temperatures detailed in Table 2) for 30 sec, and extension at 72℃ for 30 sec; and a final extension step at 72℃ for 5 min.

The Ethics Committee of the Second Hospital Affiliated to Zhejiang University approved this study, and this study was in accordance with the ethical guidelines of the Declaration of Helsinki. The experiments in this study were completely in accordance with the approved guidelines. Written informed consent was obtained from each participating individuals before sample collection.
In this study, 18 family members, including 5 affected individuals, were enrolled (Fig. 1A). Ophthalmologic examinations including uncorrected visual acuity, cycloplegic retinoscopy and manifest refractive power, best-corrected visual acuity (BCVA), ocular movements, slitlamp, and fundus examination were performed. B-scan ultrasonography (IU22, Philips) and A-scan ultrasonography (CineScan, Quantel Medical) were additionally performed for each affected patient for further evaluation.
DNA from participating family members was extracted from peripheral-blood lymphocytes by standard extraction procedures of the QuickGene DNA whole-blood kit (Fujifilm).

The genomic DNA was extracted from peripheral blood by QuickGene DNA Whole Blood Kit (Fujifilm, Tokyo, Japan) and according to the manufacturer's agreement. Genotyping was performed using iPLEX chemistry on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS, named as the MassARRAY system, which is manufactured by Sequenom, San Diego, CA). Polymerase chain reaction (PCR) and extended primers were designed using the online primer design tool Sequenom. Mass spectrum was obtained using Compact Mass Spectrometer and analyzed with MassARRAY Typer 4.0 Software.

Mutation analysis was carried out using genomic DNA from peripheral blood using a QuickGene DNA whole blood kit (Fujifilm, Tokyo, Japan). Twenty-three exons, including coding sequences of the OCRL gene and their intronic flanking sequences, were amplified by polymerase chain reaction (PCR) with 20 sets of primers. After PCR amplification with Go Taq polymerase (Promega, Madison, WI, USA), DNA sequencing was performed using the same primers as those in PCR and a BigDye Terminator V3.1 Cycle Sequencing Ready Reaction kit (Applied Biosystems, Foster City, CA, USA).